Poster 064 - Enzymatic and immunochemical characterization of B. insularis venom and serum neutralization

Poster 64. Congresso da Sociedade Brasileira de Toxinologia, 8., Symposium of the Pan American Section of the International Society on Toxinology, 8., 2004, Angra dos Reis, Brasil. Abstracts... J. Venom. Anim. Toxins incl.Trop. Dis., 2004, 10, 3, p.423.

Enzymatic And Immunochemical Characterization Of B. insularis Venom And Serum Neutralization Of The Lethal Activity By Anti-Bothropic Serum

1 Laboratory of Immunopathology; 2 Laboratory of Herpetology, Butantan Institute, SP, Brazil.

The B. insularis snake (Jararaca ilhoa) is found only at Queimada Grande Island, Brazil. Very little is known about B. insularis venom. The envenoming caused by Bothrops snake induces coagulation disturbance beside local damage like edema and hemorrhage and can result in necrosis. The aim of this work was to verify the presence of phospholipase and proteolytic activities, the antigenic cross-reactivity (ELISA and Immunoblotting) between B. insularis and B. jararaca venoms using anti-bothropic horse serum (Butantan Institute) and neutralization of lethality induced by B. insularis venom. By SDS-PAGE 12.5%, stained by silver nitrate, it was observed the existence of many common components above 50 kDa between B. jararaca and B. insularis. However, some specific antigens were observed below 42 kDa in B. jararaca venom. B. insularis venom has gelatinolytic (40 - 89 kDa and 170 kDa), caseinolytic (40 - 110 kDa and 170 kDa), fibrinogenolytic (37 - 80 kDa) and phospholipase activities. All these activities were also described in literature to B. jararaca venom. The band of 170 kDa with gelatinolytic and caseinolytic activities seems to be a metalloproteinase since the incubation with 1,10 phenantroline abolished its enzymatic activity. No hyaluronidase activity was found in these venoms. Anti-bothropic serum showed similar titer (1,280,000) to B. jararaca and B. insularis venoms by ELISA as well as recognized similar components by Immunoblotting in both venoms. B. jararaca venom is more toxic (DL50 1.63 mg/kg) than the B. insularis venom (DL50 1.98 mg/kg) to mice. The anti-bothropic serum was efficient to neutralize the lethal activity of 5 DL50 B. insularis venom. Our results showed that there is antigenic cross-reativity between B. jararaca and B. insularis venoms, suggesting that proteolytic enzymes including metalloproteinases and phospholipase found in B. insularis venom may induce local damage as observed in others Bothrops venoms.